The long-term objective of the Scientific Core is to provide outstanding, state-of-the-art, cost-effective Core facilities and services that support all five projects of the Program. Core B is composed of a protein expression and purification facility, as well as other shared facilities including a darkroom, Phosphorlmager, ultracentrifuge, and liquid nitrogen freezers. The Core will also support the generation of retroviruses expressing variou~ shRNAs and microarray analysis of miRNA expression. The Phosphorlmager will be used for imaging and quantification of various results including primerextension assays, gel mobility shifts, and Westem blots. The protein expression and purification facility consists of a shaking incubator, a centrifuge, an FPLC, and protein expression vectors. It is used to express and purify proteins that are studied in each of the projects. In a new service, the Core will also generate shRNA constructs for the various projects. This will include design of shRNAs that target specific genes, cloning the inserts into retrovirus vectors, and generation of high-titer retrovirus stocks. The Core will also support the use of a Taqman microRNA microarray set-up by purchasing cassettes for the analysis. An Amaxa Nucleofector transfection apparatus will also be purchased for use by the projects. The Core also possesses liquid nitrogen freezers for the storage of cell lines and a gel documentation system. In addition, expertise is provided to all member laboratories by personnel who oversee the Core. This Scientific Core is essential to the success of the Program, and facilitates interaction between member laboratories.